During light-adaptation by the vertebrate eye, the rods are desensitized and the light response is accelerated. When light is absorbed by the rods, a phosphodiesterase is activated that hydrolyses cyclic GMP. A light-induced decrease in cytoplasmic Ca2+ concentration is part of this light-adaptation process. The protein S-modulin (M(r) 26,000) is known to increase the fraction of light-activated cyclic GMP-phosphodiesterase (PDE) at high Ca2+ concentrations in frog rod photoreceptors. Here I present evidence that S-modulin lengthens the lifetime of active PDE (PDE*) at high Ca2+ concentrations. These S-modulin effects are observed in the physiological range of Ca2+ concentration (30 nM to 1 microM; half-maximum effects at 200-400 nM). At the high Ca2+ concentrations at which S-modulin prolongs the lifetime of PDE*, S-modulin inhibits rhodopsin phosphorylation (half-maximum effect at approximately 100 nM Ca2+). ATP is necessary for the S-modulin effects on PDE activation. I therefore conclude that the Ca(2+)-dependent regulation of PDE by S-modulin is mediated by rhodopsin phosphorylation. This regulation seems to be the principal mechanism of light adaptation in vertebrate photoreceptors.